Newly synthesized peptide, Ara-27, exhibits significant improvement in cell-penetrating ability compared to conventional peptides.


Journal

Biotechnology progress
ISSN: 1520-6033
Titre abrégé: Biotechnol Prog
Pays: United States
ID NLM: 8506292

Informations de publication

Date de publication:
09 2020
Historique:
received: 01 11 2019
revised: 17 03 2020
accepted: 21 04 2020
pubmed: 7 5 2020
medline: 29 9 2021
entrez: 7 5 2020
Statut: ppublish

Résumé

Cell-penetrating peptides (CPPs) are short amino acid sequences known to act as a vehicle for enhancing the intracellular translocating efficiency of extracellular molecules. Although many groups have attempted to develop peptides with high cell-penetrating efficiencies, very few have demonstrated efficient cellular uptake of CPPs at low concentrations. Here, we describe a newly synthesized peptide derived from Arabidopsis, Ara-27, which exhibits significant improvement in cell-penetrating efficiency compared to existing CPPs. The cell-penetrating efficiency of Ara-27 was compared with the commonly used Tat-protein transduction domain (Tat-PTD) and membrane translocating sequence (MTS) in human dermal fibroblast (HDF) and human dental pulp stem cells (hDPSC). Cell-penetrating efficiency of fluorescein isothiocyanate (FITC)-labeled CPPs were assessed by flow cytometry and visualized by confocal microscopy. Flow cytometric analysis revealed >99% cell-penetrating efficiency for 2 μM Ara-27 in both HDF and hDPSC. In contrast, 2 μM Tat-PTD and MTS showed <10% cell-penetrating efficiency in both cells. In support, relative fluorescence intensities of FITC-labeled Ara-27 were around 8 to 22 times higher than those of Tat-PTD and MTS in both cells. Confocal analysis revealed internalization of 0.2 and 2 μM Ara-27 in both human cells, which was not observed for Tat-PTD and MTS at either concentration. In conclusion, this study describes a novel CPP, Ara-27, which exhibit significant improvement in intracellular uptake compared to conventional CPPs, without affecting cell viability. Thus, development of Ara-27 based peptides may lead to improved delivery of functional cargo such as small molecules, siRNA, and drugs for in vivo studies.

Identifiants

pubmed: 32374475
doi: 10.1002/btpr.3014
doi:

Substances chimiques

Arabidopsis Proteins 0
Cell-Penetrating Peptides 0
Zinc J41CSQ7QDS

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e3014

Subventions

Organisme : National Research Foundation of Korea
ID : NRF-2016R1D1A1B03931864
Pays : International
Organisme : Technology Development Program funded by the Ministry of SMEs and Startups (MSS, Korea)
ID : S2423830
Pays : International
Organisme : Technology Development Program funded by the Ministry of SMEs and Startups (MSS, Korea)
ID : S2519744
Pays : International

Informations de copyright

© 2020 American Institute of Chemical Engineers.

Références

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Auteurs

Sol Min (S)

Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Korea.

Kichul Kim (K)

Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Korea.

Seockmo Ku (S)

Fermentation Science Program, School of Agriculture, College of Basic and Applied Sciences, Middle Tennessee State University, Murfreesboro, Tennessee, USA.

Jeong-Yoon Park (JY)

The Spine and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Republic of Korea.

Jeongmin Seo (J)

Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Korea.
Biomedical Research Institute, NeoRegen Biotech Co., Ltd., Gyeonggi-do, Korea.

Sangho Roh (S)

Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Korea.

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